Cystic Fibrosis (CF) is an autosomal recessive disease that is the result of defective CF transmembrane conductance regulator (CFTR) ion transport function at the cell surface membrane. Individuals suffering this fate can expect to live to their late-thirties. A frequent mutation affecting most CF patients is the F508del mutation found in at least one CF allele at > 80% frequency. This mutant F508del CFTR is a misfolded form of CFTR that is impaired in its ability to traffic to the cytoplasmic apical membrane where it primarily functions as a cAMP-dependent Cl channel, albeit with lowered efficiency. A long non-coding RNA (lncRNA) has been identified that is involved in transcriptional regulation of CFTR. Preliminary studies indicate that inhibition of this lncRNA can result in increased expression of mutant CFTR at the cell surface. Therefore, the studies proposed here will test the hypothesis that sustainable expression of mutant CFTR can be achieved in human cells such that there is physiologically and therapeutically efficacious CFTR-associated cAMP-dependent Cl ion transport function present at the apical membrane of epithelial cells. We have developed three aims to test our hypothesis. In aim 1, we will develop and contrast vector and oligonucleotide targeted activation of CFTR, in aim 2, we will characterize the functional components involved in lncRNA modulation of CFTR and in aim 3 we will determine the genome targeting and gene expression profile of these lncRNAs BG213071 and AI805947 in expressing and repressed cells. These studies represent the development of a new therapeutic paradigm for CF, i.e., targeting lncRNAs to affect the function of the protein-coding gene targets that they regulate. In this context, several novel approaches targeted to CFTR-associated lncRNAs will be developed and tested to enhance CFTR expression and function. This new pathway for regulation of CFTR expression by lncRNA could lead to significant insights into those cellular pathways involved in the modulation of CFTR expression during development and in response to secondary infections as well as to the development of a new class of therapeutic agents to treat CF.